Coexpression of gamma and beta globin mRNA in cells containing a single
human beta globin locus: results from studies using single-cell reverse
transcription polymerase chain reaction [published erratum appears in Blood
1994 Aug 15;84(4):1357]
T Furukawa, G Zitnik, K Leppig, T Papayannopoulou and G Stamatoyannopoulos
Department of Medicine, University of Washington, Seattle 98195.
We developed a method detecting globin gene expression in single cells
using reverse transcription polymerase chain reaction. epsilon and gamma
globin cDNAs are coamplified by an epsilon gamma primer set whereas gamma
and beta globin cDNAs are coamplified by a gamma beta primer set and the
individual globin cDNAs are distinguished by restriction enzyme digestion.
Analysis of RNA preparations from human fetal liver, neonatal red blood
cells (RBCs), or adult RBCs showed the expected mRNA species for each stage
of human development. Analysis of single cells from a human erythroleukemia
line coexpressing gamma and beta globin chains showed heterogeneity in
gamma and beta mRNA contents. The method was subsequently used to test
whether only one or more than one globin genes are expressed in cells that
contain a single human beta globin locus. We found that about 50% of single
cells from MEL x fetal erythroid cell hybrids containing a single human
beta globin locus coexpressed gamma and beta globin mRNA. This finding is
best explained by assuming that both gamma and beta genes are
simultaneously transcribed from the same beta globin locus implying that
the LCR can simultaneously interact with more than one globin gene
promoter.
Volume 83,
Issue 5,
pp. 1412-1419,
03/01/1994
Copyright © 1994 by The American Society of Hematology
